Overexpression of kin17 protein forms intranuclear foci in mammalian cells.
We used antibodies against E coli RecA protein to identify in mouse cells a 45-kDa DNA-binding protein called kin17, which has an active zinc finger and a nuclear localisation signal. Kin17 protein produced in E coli binds preferentially to the curved DNA of a bacterial promoter in vivo and in vitro, suggesting a transcriptional regulation activity. The fact that in rodent cells kin17 protein levels increase after gamma-irradiation suggests its participation in a cellular response to ionising radiation. We raised polyclonal antibodies against the whole kin17 protein and against its derived synthetic peptides. We report the detection of kin17 protein and of truncated forms of the protein by Western blot or by immunocytochemistry after transient overexpression in cultured human cells. Our results indicate that the cross-reactivity with the anti-RecA antibodies is due to an antigenic determinant located in the core of kin17 protein, between residues 129 and 228. The kin17 protein is located in the nucleus and is concentrated in small nuclear dot-like structures throughout the nucleoplasm. The RecA homologous region seems to play an essential role in the localisation of kin17 protein since the deletion of this particular region dramatically changes the form and the distribution of the intranuclear foci. We hypothesise that these dot-like structures reflect nuclear metabolism compartmentalization.
Kannouche
P
P
Département de Radiobiologie et de Radiopathologie, Centre d'Etudes de Fontenay-aux-Roses, CEA, France.
Pinon-Lataillade
G
G
Mauffrey
P
P
Faucher
C
C
Biard
D S
DS
Angulo
J F
JF
D000818:Animals; D015153:Blotting, Western; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D016254:Mutagenesis, Insertional; D016297:Mutagenesis, Site-Directed; D019913:Nuclear Localization Signals / Q000235:genetics; D009687:Nuclear Proteins; D017434:Protein Structure, Tertiary; D016601:RNA-Binding Proteins; D011938:Rec A Recombinases / Q000235:genetics; D017384:Sequence Deletion; D013347:Subcellular Fractions / Q000378:metabolism; D014162:Transfection; D016335:Zinc Fingers / Q000235:genetics
D004268:DNA-Binding Proteins; C489487:KIN protein, human; D019913:Nuclear Localization Signals; D009687:Nuclear Proteins; D016601:RNA-Binding Proteins; C071214:Kin protein, mouse; D011938:Rec A Recombinases